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World Journal of Microbiology and Biotechnology

, Volume 23, Issue 8, pp 1131–1137 | Cite as

Immobilization of Aspergillus oryzae α-galactosidase in gelatin and its application in removal of flatulence-inducing sugars in soymilk

  • K. Naganagouda
  • S. J. Prashanth
  • S. K. Shankar
  • S. K. Dhananjay
  • V. H. MulimaniEmail author
Original Paper

Abstract

Raffinose oligosaccharides (RO) are the major factors responsible for flatulence following ingestion of soybean-derived products. Removal of RO from seeds or soymilk would then have a positive impact on the acceptance of soy-based foods. In this study, α-galactosidase from Aspergillus oryzae was entrapped in gelatin using formaldehyde as the hardener. The immobilization yield was 64.3% under the optimum conditions of immobilization. The immobilized α-galactosidase showed a shift in optimum pH from 4.8 to 5.4 in acetate buffer. The optimum temperature also shifted from 50°C to 57°C compared with soluble enzyme. Immobilized α-galactosidase was used in batch, repeated batch and continuous mode to degrade RO present in soymilk. In the repeated batch, 45% reduction of RO was obtained in the fourth cycle. The performance of immobilized α-galactosidase was tested in a fluidized bed reactor at different flow rates and 86% reduction of RO in soymilk was obtained at 25 ml h−1 flow rate. The study revealed that immobilized α-galactosidase in continuous mode is efficient in reduction of RO present in soymilk.

Keywords

α-Galactosidase Aspergillus oryzae Fluidized bed reactor Gelatin Raffinose oligosaccharides Soymilk 

Notes

Acknowledgments

One of the authors Naganagouda V. Kote thanks Gulbarga University, Gulbarga for providing the financial support in the form of University Ph.D. junior research fellowship.

References

  1. Abdel-Naby MA, Sheriff AA, EI-Tanash AB, Mankarios AT (1999) Immobilization of A. oryzae tannase and properties of immobilized enzyme. J Appl Microbiol 57:108–114CrossRefGoogle Scholar
  2. Alteriis ED, Parascandola P, Salvadore S, Sardi V (1985) Enzyme immobilization within insolubilized gelatin. J Chem Technol Biotechnol 35B:60–64CrossRefGoogle Scholar
  3. Arica MY, Alaeddinoglu NG, Hasirei V (1996) Immobilization of glucoamylase onto activated PHEMA/EDGMA microspheres: properties and applications into a packed bed reactor. Enzyme Microb Technol 18:281–285CrossRefGoogle Scholar
  4. Barbasgaard, Hansen HP, Diderichsen B (1992) On the safety of Aspergillus oryzae: a review. Appl Microbiol Biotechnol 36:569–572Google Scholar
  5. Bulpin PV, Gidley MJ, Jeffcoat R, Underwood DJ (1990) Development of a biotechnological process for the modification of galactomannan polymers by with plant α-galactosidase. Carbohydr Polym 12:155–168CrossRefGoogle Scholar
  6. Cruz R, Park YK (1982) Production of fungal α-galactosidase and its application to the hydrolysis of galacto-oligosaccharides in soybean milk. J Food Sci 47:1973–1975CrossRefGoogle Scholar
  7. Dey PM, Pridham JB (1972) Biochemistry of α-galactosidase. Adv Enzymol 36:911–930Google Scholar
  8. Girigowda K, Mulimani VH (2006) Hydrolysis of galacto-oligosaccharides in soymilk by κ-carrageenan-entrapped α-galactosidase from Aspergillus oryzae. World J Microbiol Biotechnol 22:437–442CrossRefGoogle Scholar
  9. Kotwal SM, Gote MM, Khan MI, Khire JM (1999) Production, purification and characterization of a constitutive intracellular α-galactosidase from the thermophillic fungus Humicula.sp. J Ind Microbiol Biotechnol 23:661–667CrossRefGoogle Scholar
  10. LeBlanc JG, Silvestroni A, Connes C, Juillard V, Gieri GS, Piard J, Sesma F (2004) Reduction of non-digestible oligosaccharides in soymilk: application of engineered lactic acid bacteria that produce α-galactosidase. Genet Mol Res 3:432–440Google Scholar
  11. Leske KL, Jevne CJ, Coon CN (1993) Effect of oligosaccharides additions on nitrogen-corrected true metabolizable energy of soy protein concentrate. Poult Sci 72:664–668Google Scholar
  12. Mulimani VH, Ramalingam (1995) Enzymic hydrolysis of raffinose and stachyose present in soymilk by crude α-galactosidase from Gibberella fujikuroi. Biochem Mol Biol Int 36:897–905Google Scholar
  13. Munjal N, Sawhney SK (2002) Stability and properties of mushroom tyrosinase entrapped in alginate, polyacrylamide and gelatin gels. Enzyme Microb Technol 30:613–619CrossRefGoogle Scholar
  14. Palmeri G, Giardina P, Desiderio B, Morzullo L, Giamberini M, Sannia G (1994) A new enzyme immobilization procedure using copper alginate gel: application to fungal phenol oxidase. Enzyme Microb Technol 16:151–158CrossRefGoogle Scholar
  15. Porter JE, Sarikaya A, Herrmann KM, Ladisch MR (1992) Effect of pH on subunit association protection of soybean a-galactosidase. Enzyme Microb Technol 14:609–614CrossRefGoogle Scholar
  16. Prashanth SJ, Mulimani VH (2005) Soymilk oligosaccharide hydrolysis by Aspergillus oryzae α-galactosidase immobilized in calcium alginate. Process Biochem 40:1199–1205CrossRefGoogle Scholar
  17. Sahasrabudhe SR, Modi AJ, Modi VV (1988) Dehalogenation of 3-chlorobenzoate by immobilized Pseudomonas sp. B13 cells. Biotechnol Bioeng 31:889–893CrossRefGoogle Scholar
  18. Scalabrini P, Rossi M, Spettoli P, Matteuzzi D (1998) Characterization of Bifidobacterium strains for use in soymilk fermentation. Int J Food Microbiol 39:213–219CrossRefGoogle Scholar
  19. Scordi V (1987) Immobilization of enzyme and microbial cells in gelatin. Meth Enzymol 135:293–299CrossRefGoogle Scholar
  20. Slominski BA (1994) Hydrolysis of galactooligosaccharides by commercial preparations of α-galactosidase and β-fructofuranose: potential for use as dietary additives. J Agric Food Chem 65:323–330CrossRefGoogle Scholar
  21. Song D, Chang KC (2006) Enzymatic degradation of oligosaccharides in pinto bean flour. J Agric Food Chem 54:1296–1301CrossRefGoogle Scholar
  22. Srivastava PK, Kayastha AM, Srinivasan (2001) Characterization of gelatin-immobilized pigeonpea urease and preparation of a new urea biosensor. Biotechnol Appl Biochem 34:55–62CrossRefGoogle Scholar
  23. Steggerda FR, Richard EA, Rackis JJ (1966) Effects of various soybean products on flatulence in the adult man. Proc Soc Exp Biol Med 121:1235–1239Google Scholar
  24. Suarez FL, Springfield J, Furne JK, Lohrmann TT, Kerr PS, Levitt MD (1999) Gas production in humans ingesting a soybean flour derived from beans naturally low in oligosaccharides. Am J Clin Nutr 69:135–139Google Scholar
  25. Sungur S, Akbulut V (1994) Immobilization of β-galactosidase on to gelatin by glutaraldehyde and chromium (III) acetate. J Chem Technol Biotechnol 59:303–306CrossRefGoogle Scholar
  26. Tanaka M, Thananunkul D, Lee TC, Chichester CO (1975) A simplified method for the quantitative determination of sucrose, raffinose and stachyose in legumes. J Food Sci 40:1087–1088CrossRefGoogle Scholar
  27. Thananunkul D, Tanaka M, Chichester CO, Lee TC (1976) Degradation of raffinose and stachyose in soybean milk by α-galactosidase from Mortierella vinacea. Entrapment of α-galactosidase within polyacrylamide gel. J Food Sci 41:173–175CrossRefGoogle Scholar
  28. Thippeswamy S, Mulimani VH (2002) Enzymic degradation of raffinose family oligosaccharides in soymilk by immobilized α-galactosidase from Gibberella fujikuroi. Process Biochem 38:635–640CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • K. Naganagouda
    • 1
  • S. J. Prashanth
    • 2
  • S. K. Shankar
    • 1
  • S. K. Dhananjay
    • 1
  • V. H. Mulimani
    • 1
    Email author
  1. 1.Department of Biochemistry Gulbarga UniversityGulbargaIndia
  2. 2.Department of Biochemistry Indian Institute of ScienceBangaloreIndia

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